A molecular assay for sensitive detection of pathogen-specific T-cells.
- Kasprowicz, Victoria O
- Mitchell, Jessica E
- Chetty, Shivan
- Govender, Pamla
- Huang, Kuan-Hsiang Gary
- Fletcher, Helen A
- Webster, Daniel P
- Brown, Sebastian
- Kasmar, Anne
- Millington, Kerry
- Day, Cheryl L
- Mkhwanazi, Nompumelelo
- McClurg, Cheryl
- Chonco, Fundisiwe
- Lalvani, Ajit
- Walker, Bruce D
- Ndung'u, Thumbi
- Klenerman, Paul
DOIAbstract
Here we describe the development and validation of a highly sensitive assay of antigen-specific IFN-γ production using real time quantitative PCR (qPCR) for two reporters--monokine-induced by IFN-γ (MIG) and the IFN-γ inducible protein-10 (IP10). We developed and validated the assay and applied it to the detection of CMV, HIV and Mycobacterium tuberculosis (MTB) specific responses, in a cohort of HIV co-infected patients. We compared the sensitivity of this assay to that of the ex vivo RD1 (ESAT-6 and CFP-10)-specific IFN-γ Elispot assay. We observed a clear quantitative correlation between the two assays (P<0.001). Our assay proved to be a sensitive assay for the detection of MTB-specific T cells, could be performed on whole blood samples ...
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